Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Isolation

Human, Dakar, 1950

Product Format

frozen

Storage Conditions

Frozen Cultures:-70°C for 1 week; liquid N2 vapor for long term storage

Harvest cells from a culture which is at or near peak density by centrifugation at 1,300 g for 5 min.

Adjust concentration of cells to 2 x 107/mL in fresh medium.

While cells are centrifuging prepare a 10% (v/v) solution of sterile DMSO in fresh medium (broth). The DMSO solution when first prepared will warm up due to chemical heat. The solution should be allowed to return to room temperature prior to use.

Mix the cell preparation and the DMSO solution in equal portions. The final concentration will be 107 cells/mL and 5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no more than 15 min.

Place the ampules in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)

Store in either the vapor or liquid phase of a nitrogen refrigerator.

To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes. Do not agitate the ampule. Do not leave ampule in water bath after thawed.

Immediately after thawing, do not leave in the water bath, aseptically transfer the contents of the ampule into a fresh flask containing 10 mL ATCC Medium 2831.

Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.